PD-1/PD-L1 blockade cell-based assay

Current methods to measure the potency of drugs targeting PD-1 or PD-L1 include in vitro binding assays, primary T cell-based cytokine release assays, or in vivo model systems. To complete our in vivo services, we also offer a robust, quantitative, functional bioassay to identify and characterize in 96-well plates the potency of candidate drugs capable of modulating the interaction of PD-1/PD-L1.

Assay Principle

  • Artificial Antigen Presenting cells (CHO) expressing at their surface MHC and PDL1 are plated in 96-well plates with effector T Cells (Jurkat) engineered to present PD1 and to express a bioluminescent reporter gene under the control of the TCR signaling pathway.
  • T Cell Receptor engagement induces luciferase activity in the effector T cell (Jurkat) while co-engagement of an immune checkpoint receptor-ligand pair inhibits luciferase activity
  • Antibody-mediated blockade of the immune checkpoint inhibitory signal restores luciferase activity that is detected using appropriated luminescent plate reader

anti pd1pdl1 cell-based assay immune checkpoint inhibitor screening CRO service cancer immunotherapy

Representative Results

In vitro assay_antiPD1_PDL1

PD-1 expressing Effector Cells and PD-L1 APC cells were incubated for 6 hrs at 37°C with increasing concentrations of either an anti-PD-1, PD-L1, or CTLA-4 antibodies. The anti-PD-1 and PD-L1, but not the anti-CTLA-4 antibody, abolishes the immune checkpoint inhibitory signal resulting in the restoration of luciferase activity.

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Explicyte immuno-oncology offers to test novel cancer immunotherapies for their ability to induce PD-1/PD-L1 blockade. Our team has validated an anti-PD-1/PD-L1 cell-based assay for the screening of novel inhibitors.